title: 'Module'-substituted globins: artificial exon shuffling among myoglobin, hemoglobin alpha- and beta-subunits.
abstract: Based on the detailed structural analysis of proteins, Go [M. Go, Nature 291 (1981) 90-92] found that protein structures can be divided into some structural units, 'modules,' which correspond to peptides coded by exons. In the present study, to investigate functional and structural roles of modular structures in proteins, we have engineered eight chimera globins, in which the exons are shuffled among human myoglobin, human hemoglobin alpha- and beta-subunits, in addition to the chimera beta beta alpha-globin described previously [K. Wakasugi, K. Ishimori, K. Imai, Y. Wada, I. Morishima, J. Biol. Chem. 269 (1994) 18750-18756]. Although all of the chimera globins stoichiometrically bound the heme and their alpha-helical contents increased by heme incorporation as found for native globins, the alpha-helical contents of the chimera globins were significantly lower than those of native globins, suggesting that 'module' substitutions seriously affect the protein folding and stability in globins. The comparisons among several chimera globins demonstrated that such structural alterations are mainly attributed to loss of some key intermodular interactions for protein folding. By simultaneous substitution of the modules M1 and M4 from the same globin, the protein structure was stabilized, which indicates that the module packing between modules M1 and M4 would be one of the crucial interaction to stabilize the globin fold. Present results allow us to conclude that module substitutions would be available for designing and producing novel functional proteins if we can reproduce the stable modular packing in the 'module'-substituted proteins.
authors.affiliation: Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Wakasugi
authors.forename: K
authors.initials: K
authors.lastname: Ishimori
authors.forename: I
authors.initials: I
authors.lastname: Morishima
mesh_terms: D002850:Chromatography, Gel; D002942:Circular Dichroism; D005091:Exons; D006454:Hemoglobins / Q000235:genetics*; D006801:Humans; D046911:Macromolecular Substances; D009211:Myoglobin / Q000235:genetics*; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
chemical_list: D006454:Hemoglobins; D046911:Macromolecular Substances; D009211:Myoglobin; D011993:Recombinant Fusion Proteins